Ethanol tolerance in engineered strains of Clostridium thermocellum

Olson, Daniel G.; Maloney, Marybeth I.; Lanahan, Anthony A.; Cervenka, Nicholas D.; Xia, Ying; Pech-Canul, Angel; Hon, Shuen; Tian, Liang; Ziegler, Samantha J.; Bomble, Yannick J.; Lynd, Lee R.

doi:10.1186/s13068-023-02379-z

Ethanol tolerance in engineered strains of Clostridium thermocellum

Journal Article · Thu Sep 14 00:00:00 EDT 2023 · Biotechnology for Biofuels and Bioproducts

DOI:https://doi.org/10.1186/s13068-023-02379-z· OSTI ID:2000112

Olson, Daniel G.; Maloney, Marybeth I.; Lanahan, Anthony A.; Cervenka, Nicholas D.; Xia, Ying; Pech-Canul, Angel; Hon, Shuen; Tian, Liang; Ziegler, Samantha J.; Bomble, Yannick J.; Lynd, Lee R.

Abstract

Clostridium thermocellum is a natively cellulolytic bacterium that is promising candidate for cellulosic biofuel production, and can produce ethanol at high yields (75–80% of theoretical) but the ethanol titers produced thus far are too low for commercial application. In several strains of C. thermocellum engineered for increased ethanol yield, ethanol titer seems to be limited by ethanol tolerance. Previous work to improve ethanol tolerance has focused on the WT organism. In this work, we focused on understanding ethanol tolerance in several engineered strains of C. thermocellum . We observed a tradeoff between ethanol tolerance and production. Adaptation for increased ethanol tolerance decreases ethanol production. Second, we observed a consistent genetic response to ethanol stress involving mutations at the AdhE locus. These mutations typically reduced NADH-linked ADH activity. About half of the ethanol tolerance phenotype could be attributed to the elimination of NADH-linked activity based on a targeted deletion of adhE . Finally, we observed that rich growth medium increases ethanol tolerance, but this effect is eliminated in an adhE deletion strain. Together, these suggest that ethanol inhibits growth and metabolism via a redox-imbalance mechanism. The improved understanding of mechanisms of ethanol tolerance described here lays a foundation for developing strains of C. thermocellum with improved ethanol production.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities (SUF); USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231; AC05-00OR22725; AC36-08GO28308

OSTI ID:: 2000112

Alternate ID(s):: OSTI ID: 2203231

Report Number(s):: NREL/JA--2700-87818; 137; PII: 2379

Journal Information:: Biotechnology for Biofuels and Bioproducts, Journal Name: Biotechnology for Biofuels and Bioproducts Journal Issue: 1 Vol. 16; ISSN 2731-3654

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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